resolution standard for MP and SHG?

[Cromey, Douglas W - (dcromey)]

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I am collaborating with a group that is building a multiphoton microscope. Can anyone recommend resolution test samples for 2P and second/third harmonic imaging? Pictures are nice, but we would like to test performance as well.
Doug

------------------------------------------------------------------------------------------
Douglas W. Cromey, M.S. - Associate Scientific Investigator
Dept. of Cellular & Molecular Medicine, University of Arizona
1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA

office:  UACC 0914A       email: [hidden email]<mailto:[hidden email]>
voice:  520-626-2824       fax:  [phone]

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Home of: "Microscopy and Imaging Resources on the WWW"

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[Katrin Roth-2]

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Dear Doug,

our application specialist from Olympus told me to use fluorescent beads for quality management.  The 1-2µm beads for finding the right planar  and 500nm  beads for measuring (the shape). Perhaps you can use even smaller beads up to resolution frontiers.

All the best
Katrin


Am 13.11.2015 um 15:42 schrieb "Cromey, Douglas W - (dcromey)" <[hidden email]>:

> *****
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> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> I am collaborating with a group that is building a multiphoton microscope. Can anyone recommend resolution test samples for 2P and second/third harmonic imaging? Pictures are nice, but we would like to test performance as well.
> Doug
>
> ------------------------------------------------------------------------------------------
> Douglas W. Cromey, M.S. - Associate Scientific Investigator
> Dept. of Cellular & Molecular Medicine, University of Arizona
> 1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA
>
> office:  UACC 0914A       email: [hidden email]<mailto:[hidden email]>
> voice:  520-626-2824       fax:  [phone]
>
> http://swehsc.pharmacy.arizona.edu/micro
> Home of: "Microscopy and Imaging Resources on the WWW"
>
> UA Microscopy Alliance -  http://microscopy.arizona.edu<http://microscopy.arizona.edu/>
>

––––––––––––––––––––––––––––––––
Dr. Katrin Roth
Philipps-University of Marburg
Core Facility Cellular Imaging in
Centre of cancer and immunology (ZTI)
Hans-Meerweinstr. 3
35043 Marburg
Germany

Tel: [phone]

[Kurt Anderson-3]

dear doug,

urea crystals are a good sample if you just want to know whether you can generate any SHG at all, but they don’t have much structural detail and aren’t very biologically relevant. collagen is an excellent second harmonic generator, and one of the biological materials you are likely to image in the end. you should be able to fix a bit of tissue and keep that around, although fresh material might have better ultrastructure. there are many excellent sources:

there is a dense layer of collagen in the mouse intestine between the smooth muscle and base of the intestinal crypt. the structural detail is very rich, and you should be able to resolve very thin fibrils within larger bundles. this one is my personal favourite; if you contact me off-list i can send you images.

alternatively, you might image rat tail tendons:
Biophys J.<http://www.ncbi.nlm.nih.gov/pubmed/15533922#> 2005 Feb;88(2):1377-86. Epub 2004 Nov 8.
Interpreting second-harmonic generation images of collagen I fibrils.
Williams RM<http://www.ncbi.nlm.nih.gov/pubmed/?term=Williams%20RM%5BAuthor%5D&cauthor=true&cauthor_uid=15533922>1, Zipfel WR<http://www.ncbi.nlm.nih.gov/pubmed/?term=Zipfel%20WR%5BAuthor%5D&cauthor=true&cauthor_uid=15533922>, Webb WW<http://www.ncbi.nlm.nih.gov/pubmed/?term=Webb%20WW%5BAuthor%5D&cauthor=true&cauthor_uid=15533922>.

or skin, e.g.:
Cancer Cell.<http://www.ncbi.nlm.nih.gov/pubmed/?term=Samuel%2C+Olson%2C+anderson%2C+mcghee#> 2011 Jun 14;19(6):776-91. doi: 10.1016/j.ccr.2011.05.008.
Actomyosin-mediated cellular tension drives increased tissue stiffness and β-catenin activation to induce epidermal hyperplasia and tumor growth.
Samuel MS<http://www.ncbi.nlm.nih.gov/pubmed/?term=Samuel%20MS%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>1, Lopez JI<http://www.ncbi.nlm.nih.gov/pubmed/?term=Lopez%20JI%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, McGhee EJ<http://www.ncbi.nlm.nih.gov/pubmed/?term=McGhee%20EJ%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Croft DR<http://www.ncbi.nlm.nih.gov/pubmed/?term=Croft%20DR%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Strachan D<http://www.ncbi.nlm.nih.gov/pubmed/?term=Strachan%20D%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Timpson P<http://www.ncbi.nlm.nih.gov/pubmed/?term=Timpson%20P%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Munro J<http://www.ncbi.nlm.nih.gov/pubmed/?term=Munro%20J%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Schröder E<http://www.ncbi.nlm.nih.gov/pubmed/?term=Schr%C3%B6der%20E%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Zhou J<http://www.ncbi.nlm.nih.gov/pubmed/?term=Zhou%20J%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Brunton VG<http://www.ncbi.nlm.nih.gov/pubmed/?term=Brunton%20VG%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Barker N<http://www.ncbi.nlm.nih.gov/pubmed/?term=Barker%20N%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Clevers H<http://www.ncbi.nlm.nih.gov/pubmed/?term=Clevers%20H%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Sansom OJ<http://www.ncbi.nlm.nih.gov/pubmed/?term=Sansom%20OJ%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Anderson KI<http://www.ncbi.nlm.nih.gov/pubmed/?term=Anderson%20KI%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>,Weaver VM<http://www.ncbi.nlm.nih.gov/pubmed/?term=Weaver%20VM%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Olson MF<http://www.ncbi.nlm.nih.gov/pubmed/?term=Olson%20MF%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>.

have fun,

kurt

.



Prof. Kurt I. Anderson
Tumor Cell Migration Lab and
Beatson Advanced Imaging Resource (BAIR)
The Beatson Institute for Cancer Research
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On 13 Nov 2015, at 14:42, Cromey, Douglas W - (dcromey) <[hidden email]<mailto:[hidden email]>> wrote:

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I am collaborating with a group that is building a multiphoton microscope. Can anyone recommend resolution test samples for 2P and second/third harmonic imaging? Pictures are nice, but we would like to test performance as well.
Doug

------------------------------------------------------------------------------------------
Douglas W. Cromey, M.S. - Associate Scientific Investigator
Dept. of Cellular & Molecular Medicine, University of Arizona
1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA

office:  UACC 0914A       email: [hidden email]<mailto:[hidden email]>
voice:  520-626-2824       fax:  [phone]

http://swehsc.pharmacy.arizona.edu/micro
Home of: "Microscopy and Imaging Resources on the WWW"

UA Microscopy Alliance -  http://microscopy.arizona.edu<http://microscopy.arizona.edu/>

[Michael Giacomelli]

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Hi Doug,

The bead method is the standard way.  Just be aware that theres a
rather annoying tradeoff between light emission and accuracy of the
PSF measurement, with tiny beads giving you a nicer PSF but also being
a lot harder to find/see (particularly if your PSF or scanners are not
optimal).

You can also buy beads that are pre-distributed on glass slide, but
the ones I've ordered tended to be quite sparse.  These are probably
meant for a 10+ fps commercial microscope with low latency preview,
but can be really painful on a home built system, particularly if
you're still working on the software and don't have a well optimized
preview/scanning routine debugged yet.  In that case I'd recommend
putting a lot of beads on a coverslip, or even sonicating them into
PDMS so that you have less trouble finding them.

Mike

On Fri, Nov 13, 2015 at 9:58 AM, Katrin Roth
<[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Doug,
>
> our application specialist from Olympus told me to use fluorescent beads for quality management.  The 1-2µm beads for finding the right planar  and 500nm  beads for measuring (the shape). Perhaps you can use even smaller beads up to resolution frontiers.
>
> All the best
> Katrin
>
>
> Am 13.11.2015 um 15:42 schrieb "Cromey, Douglas W - (dcromey)" <[hidden email]>:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> I am collaborating with a group that is building a multiphoton microscope. Can anyone recommend resolution test samples for 2P and second/third harmonic imaging? Pictures are nice, but we would like to test performance as well.
>> Doug
>>
>> ------------------------------------------------------------------------------------------
>> Douglas W. Cromey, M.S. - Associate Scientific Investigator
>> Dept. of Cellular & Molecular Medicine, University of Arizona
>> 1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA
>>
>> office:  UACC 0914A       email: [hidden email]<mailto:[hidden email]>
>> voice:  520-626-2824       fax:  [phone]
>>
>> http://swehsc.pharmacy.arizona.edu/micro
>> Home of: "Microscopy and Imaging Resources on the WWW"
>>
>> UA Microscopy Alliance -  http://microscopy.arizona.edu<http://microscopy.arizona.edu/>
>>
>
>
>
> ––––––––––––––––––––––––––––––––
> Dr. Katrin Roth
> Philipps-University of Marburg
> Core Facility Cellular Imaging in
> Centre of cancer and immunology (ZTI)
> Hans-Meerweinstr. 3
> 35043 Marburg
> Germany
>
> Tel: [phone]

[Gautier Papon]

Dear Doug,

I would also like to point out that Argolight slide are compatible with MP and exhibit SHG contrast on their patterns. The only thing is that you must be extremely careful to use the minimal amount of energy possible with MP on the slide, as you the 100 GW/CM^2 limit is for instantaneous irradiance no averaged one.

Best regards,
Gautier


Dr. Gautier PAPON
CEO – Argolight

------------------------------------
Tel : [phone]
Argolight, A Precision Company
Www.argolight.com<http://www.argolight.com/>
——————————
Argolight is Carl Zeiss Choice for Calibration & monitoring – Read About it on www.argolight.com

De : Confocal Microscopy List au nom de Kurt Anderson
Répondre à : Confocal Microscopy List
Date : vendredi 13 novembre 2015 16:58
À : "[hidden email]<mailto:[hidden email]>"
Objet : Re: resolution standard for MP and SHG?


dear doug,

urea crystals are a good sample if you just want to know whether you can generate any SHG at all, but they don’t have much structural detail and aren’t very biologically relevant. collagen is an excellent second harmonic generator, and one of the biological materials you are likely to image in the end. you should be able to fix a bit of tissue and keep that around, although fresh material might have better ultrastructure. there are many excellent sources:

there is a dense layer of collagen in the mouse intestine between the smooth muscle and base of the intestinal crypt. the structural detail is very rich, and you should be able to resolve very thin fibrils within larger bundles. this one is my personal favourite; if you contact me off-list i can send you images.

alternatively, you might image rat tail tendons:
Biophys J.<http://www.ncbi.nlm.nih.gov/pubmed/15533922#> 2005 Feb;88(2):1377-86. Epub 2004 Nov 8.
Interpreting second-harmonic generation images of collagen I fibrils.
Williams RM<http://www.ncbi.nlm.nih.gov/pubmed/?term=Williams%20RM%5BAuthor%5D&cauthor=true&cauthor_uid=15533922>1, Zipfel WR<http://www.ncbi.nlm.nih.gov/pubmed/?term=Zipfel%20WR%5BAuthor%5D&cauthor=true&cauthor_uid=15533922>, Webb WW<http://www.ncbi.nlm.nih.gov/pubmed/?term=Webb%20WW%5BAuthor%5D&cauthor=true&cauthor_uid=15533922>.

or skin, e.g.:
Cancer Cell.<http://www.ncbi.nlm.nih.gov/pubmed/?term=Samuel%2C+Olson%2C+anderson%2C+mcghee#> 2011 Jun 14;19(6):776-91. doi: 10.1016/j.ccr.2011.05.008.
Actomyosin-mediated cellular tension drives increased tissue stiffness and β-catenin activation to induce epidermal hyperplasia and tumor growth.
Samuel MS<http://www.ncbi.nlm.nih.gov/pubmed/?term=Samuel%20MS%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>1, Lopez JI<http://www.ncbi.nlm.nih.gov/pubmed/?term=Lopez%20JI%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, McGhee EJ<http://www.ncbi.nlm.nih.gov/pubmed/?term=McGhee%20EJ%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Croft DR<http://www.ncbi.nlm.nih.gov/pubmed/?term=Croft%20DR%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Strachan D<http://www.ncbi.nlm.nih.gov/pubmed/?term=Strachan%20D%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Timpson P<http://www.ncbi.nlm.nih.gov/pubmed/?term=Timpson%20P%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Munro J<http://www.ncbi.nlm.nih.gov/pubmed/?term=Munro%20J%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Schröder E<http://www.ncbi.nlm.nih.gov/pubmed/?term=Schr%C3%B6der%20E%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Zhou J<http://www.ncbi.nlm.nih.gov/pubmed/?term=Zhou%20J%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Brunton VG<http://www.ncbi.nlm.nih.gov/pubmed/?term=Brunton%20VG%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Barker N<http://www.ncbi.nlm.nih.gov/pubmed/?term=Barker%20N%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Clevers H<http://www.ncbi.nlm.nih.gov/pubmed/?term=Clevers%20H%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Sansom OJ<http://www.ncbi.nlm.nih.gov/pubmed/?term=Sansom%20OJ%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Anderson KI<http://www.ncbi.nlm.nih.gov/pubmed/?term=Anderson%20KI%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>,Weaver VM<http://www.ncbi.nlm.nih.gov/pubmed/?term=Weaver%20VM%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>, Olson MF<http://www.ncbi.nlm.nih.gov/pubmed/?term=Olson%20MF%5BAuthor%5D&cauthor=true&cauthor_uid=21665151>.

have fun,

kurt

.



Prof. Kurt I. Anderson
Tumor Cell Migration Lab and
Beatson Advanced Imaging Resource (BAIR)
The Beatson Institute for Cancer Research
Garscube Estate, Switchback Road, Glasgow  G61 1BD



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On 13 Nov 2015, at 14:42, Cromey, Douglas W - (dcromey) <[hidden email]<mailto:[hidden email]><mailto:[hidden email]>> wrote:

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I am collaborating with a group that is building a multiphoton microscope. Can anyone recommend resolution test samples for 2P and second/third harmonic imaging? Pictures are nice, but we would like to test performance as well.
Doug

------------------------------------------------------------------------------------------
Douglas W. Cromey, M.S. - Associate Scientific Investigator
Dept. of Cellular & Molecular Medicine, University of Arizona
1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA

office:  UACC 0914A       email: [hidden email]<mailto:[hidden email]><mailto:[hidden email]>
voice:  520-626-2824       fax:  [phone]

http://swehsc.pharmacy.arizona.edu/micro
Home of: "Microscopy and Imaging Resources on the WWW"

UA Microscopy Alliance -  http://microscopy.arizona.edu<http://microscopy.arizona.edu/>

[Steffen Dietzel]

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Doug,

for THG resolution we used nanoparticles, eg. TiO2. We have tried
sunscreen, but it didn't work, too much THG signals from the fat
droplets. So pure particles are needed, attached to a slide or in
agarose or the like. Zinc oxide nanoparticles have been described to
generate SHG*. I would expect them to also produce THG, so you might get
two birds with one stone.

For z-resolution in THG, a glass/water interface should do: With a
dipping water objective, immersion water directly on a glass slide
(previously marked with a black dot to find the focal plane in bright
field or fluorescence). This sample is my favorite to check illumination
intensity across the field of view.

If you should go for the collagen fibers mentioned by Kurt, in addition
to the SHG from the fiber core, you'll get a THG signal from the
fiber/surroundings interface. At high resolution and with a sufficiently
thick fiber the THG signal is hollow at the core, were the SHG signal
is. I am not sure though how well they will do for determining
resolution, my impression was that some may be pretty thick.

Steffen

*For example: Urban, B.E., P.B. Neogi, S.J. Butler, Y. Fujita, and A.
Neogi. 2012. Second harmonic imaging of plants tissues and cell
implosion using two-photon process in ZnO nanoparticles. Journal of
biophotonics. 5:283-291.

Am 13.11.2015 um 15:42 schrieb Cromey, Douglas W - (dcromey):

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images onhttp://www.imgur.com  and include the link in your posting.
> *****
>
> I am collaborating with a group that is building a multiphoton microscope. Can anyone recommend resolution test samples for 2P and second/third harmonic imaging? Pictures are nice, but we would like to test performance as well.
> Doug
>
> ------------------------------------------------------------------------------------------
> Douglas W. Cromey, M.S. - Associate Scientific Investigator
> Dept. of Cellular & Molecular Medicine, University of Arizona
> 1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA
>
> office:  UACC 0914A       email:[hidden email]<mailto:[hidden email]>
> voice:  520-626-2824       fax:  [phone]
>
> http://swehsc.pharmacy.arizona.edu/micro
> Home of: "Microscopy and Imaging Resources on the WWW"
>
> UA Microscopy Alliance -http://microscopy.arizona.edu<http://microscopy.arizona.edu/>
>

--
------------------------------------------------------------
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Ludwig-Maximilians-Universität München
Biomedical Center (BMC)
Head of the Core Facility Bioimaging

Großhaderner Straße 9
D-82152 Planegg-Martinsried
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[Andreas Bruckbauer]

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The PS Speck microspheres work quite well, they are bright enough for 2 photon excited fluorescence. With 170 nm they are a bit big to be point sources, but I found them to be a good compromise.

Andreas

-----Original Message-----
From: "Cromey, Douglas W - (dcromey)" <[hidden email]>
Sent: ‎13/‎11/‎2015 14:42
To: "[hidden email]" <[hidden email]>
Subject: resolution standard for MP and SHG?

*****
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Post images on http://www.imgur.com and include the link in your posting.
*****

I am collaborating with a group that is building a multiphoton microscope. Can anyone recommend resolution test samples for 2P and second/third harmonic imaging? Pictures are nice, but we would like to test performance as well.
Doug

------------------------------------------------------------------------------------------
Douglas W. Cromey, M.S. - Associate Scientific Investigator
Dept. of Cellular & Molecular Medicine, University of Arizona
1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA

office:  UACC 0914A       email: [hidden email]<mailto:[hidden email]>
voice:  520-626-2824       fax:  [phone]

http://swehsc.pharmacy.arizona.edu/micro
Home of: "Microscopy and Imaging Resources on the WWW"

UA Microscopy Alliance -  http://microscopy.arizona.edu<http://microscopy.arizona.edu/>

[Sivaguru, Mayandi]

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Doug, one of the best standard for SHG and checking your polarization
state of your fundamental is starch. Grab a potato from your pantry,
take a small piece and liberate starch grains between two cover glass in
a drop of water, you are good to go. You can keep that tuber in your
refrigerator for at least a month or two.
Shiv

On 11/13/2015 8:42 AM, Cromey, Douglas W - (dcromey) wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> I am collaborating with a group that is building a multiphoton microscope. Can anyone recommend resolution test samples for 2P and second/third harmonic imaging? Pictures are nice, but we would like to test performance as well.
> Doug
>
> ------------------------------------------------------------------------------------------
> Douglas W. Cromey, M.S. - Associate Scientific Investigator
> Dept. of Cellular & Molecular Medicine, University of Arizona
> 1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA
>
> office:  UACC 0914A       email: [hidden email]<mailto:[hidden email]>
> voice:  520-626-2824       fax:  [phone]
>
> http://swehsc.pharmacy.arizona.edu/micro
> Home of: "Microscopy and Imaging Resources on the WWW"
>
> UA Microscopy Alliance -  http://microscopy.arizona.edu<http://microscopy.arizona.edu/>

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